dharmafect-1 Search Results


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  • 99
    Thermo Fisher agent dharmafect 1
    Agent Dharmafect 1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GE Healthcare dharmafect 1
    Dharmafect 1, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 92/100, based on 142 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Horizon Discovery dharmafect 1
    ( a ) A stably transfected vector in HeLa cells expresses two luciferase genes. Firefly luciferase is expressed constitutively, whereas expression of the renilla luciferase gene is under the control of miR-15b as part of the RNA-induced silencing complex (RISC). In the presence of an anti-miR-15b oligonucleotide (TP ODN in this figure), the RISC dissociates from the renilla gene. This leads to an increase in renilla luciferase activity. Comparison of the activity of renilla luciferase to the unregulated firefly luciferase yields an indication of the activity of an oligonucleotide as an anti-miR-15b oligonucleotide. ( a ) Adapted from the Integrated DNA Technologies Inc. (Coralville, IA, USA) website. The stably transfected HeLa cell line and assay protocols were developed by miRagen Therapeutics, Inc. ( b ) Lipid transfection using <t>DharmaFECT</t> 1 (Dharmacon). ( c ) Passive transfection using TP ODNs. Positive and negative controls were purchased from Dharmacon: miRIDIAN Hairpin Inhibitor (IH-300587-07) and miRIDIAN Hairpin Inhibitor Negative Control #1 (IN-001005-01).
    Dharmafect 1, supplied by Horizon Discovery, used in various techniques. Bioz Stars score: 93/100, based on 2200 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher dharmafect 1
    ( a ) A stably transfected vector in HeLa cells expresses two luciferase genes. Firefly luciferase is expressed constitutively, whereas expression of the renilla luciferase gene is under the control of miR-15b as part of the RNA-induced silencing complex (RISC). In the presence of an anti-miR-15b oligonucleotide (TP ODN in this figure), the RISC dissociates from the renilla gene. This leads to an increase in renilla luciferase activity. Comparison of the activity of renilla luciferase to the unregulated firefly luciferase yields an indication of the activity of an oligonucleotide as an anti-miR-15b oligonucleotide. ( a ) Adapted from the Integrated DNA Technologies Inc. (Coralville, IA, USA) website. The stably transfected HeLa cell line and assay protocols were developed by miRagen Therapeutics, Inc. ( b ) Lipid transfection using <t>DharmaFECT</t> 1 (Dharmacon). ( c ) Passive transfection using TP ODNs. Positive and negative controls were purchased from Dharmacon: miRIDIAN Hairpin Inhibitor (IH-300587-07) and miRIDIAN Hairpin Inhibitor Negative Control #1 (IN-001005-01).
    Dharmafect 1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 1486 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Fisher Scientific dharmafect 1
    ( a ) A stably transfected vector in HeLa cells expresses two luciferase genes. Firefly luciferase is expressed constitutively, whereas expression of the renilla luciferase gene is under the control of miR-15b as part of the RNA-induced silencing complex (RISC). In the presence of an anti-miR-15b oligonucleotide (TP ODN in this figure), the RISC dissociates from the renilla gene. This leads to an increase in renilla luciferase activity. Comparison of the activity of renilla luciferase to the unregulated firefly luciferase yields an indication of the activity of an oligonucleotide as an anti-miR-15b oligonucleotide. ( a ) Adapted from the Integrated DNA Technologies Inc. (Coralville, IA, USA) website. The stably transfected HeLa cell line and assay protocols were developed by miRagen Therapeutics, Inc. ( b ) Lipid transfection using <t>DharmaFECT</t> 1 (Dharmacon). ( c ) Passive transfection using TP ODNs. Positive and negative controls were purchased from Dharmacon: miRIDIAN Hairpin Inhibitor (IH-300587-07) and miRIDIAN Hairpin Inhibitor Negative Control #1 (IN-001005-01).
    Dharmafect 1, supplied by Fisher Scientific, used in various techniques. Bioz Stars score: 92/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche dharmafect 1
    ( a ) A stably transfected vector in HeLa cells expresses two luciferase genes. Firefly luciferase is expressed constitutively, whereas expression of the renilla luciferase gene is under the control of miR-15b as part of the RNA-induced silencing complex (RISC). In the presence of an anti-miR-15b oligonucleotide (TP ODN in this figure), the RISC dissociates from the renilla gene. This leads to an increase in renilla luciferase activity. Comparison of the activity of renilla luciferase to the unregulated firefly luciferase yields an indication of the activity of an oligonucleotide as an anti-miR-15b oligonucleotide. ( a ) Adapted from the Integrated DNA Technologies Inc. (Coralville, IA, USA) website. The stably transfected HeLa cell line and assay protocols were developed by miRagen Therapeutics, Inc. ( b ) Lipid transfection using <t>DharmaFECT</t> 1 (Dharmacon). ( c ) Passive transfection using TP ODNs. Positive and negative controls were purchased from Dharmacon: miRIDIAN Hairpin Inhibitor (IH-300587-07) and miRIDIAN Hairpin Inhibitor Negative Control #1 (IN-001005-01).
    Dharmafect 1, supplied by Roche, used in various techniques. Bioz Stars score: 92/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GE Healthcare dharmafect 1 reagent
    ( a ) A stably transfected vector in HeLa cells expresses two luciferase genes. Firefly luciferase is expressed constitutively, whereas expression of the renilla luciferase gene is under the control of miR-15b as part of the RNA-induced silencing complex (RISC). In the presence of an anti-miR-15b oligonucleotide (TP ODN in this figure), the RISC dissociates from the renilla gene. This leads to an increase in renilla luciferase activity. Comparison of the activity of renilla luciferase to the unregulated firefly luciferase yields an indication of the activity of an oligonucleotide as an anti-miR-15b oligonucleotide. ( a ) Adapted from the Integrated DNA Technologies Inc. (Coralville, IA, USA) website. The stably transfected HeLa cell line and assay protocols were developed by miRagen Therapeutics, Inc. ( b ) Lipid transfection using <t>DharmaFECT</t> 1 (Dharmacon). ( c ) Passive transfection using TP ODNs. Positive and negative controls were purchased from Dharmacon: miRIDIAN Hairpin Inhibitor (IH-300587-07) and miRIDIAN Hairpin Inhibitor Negative Control #1 (IN-001005-01).
    Dharmafect 1 Reagent, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 90/100, based on 39 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher dharmafect 1 reagent
    Suppressive effect of novel RNAi agents for RPN2 in A549-luc-C8 cells. (a) Immunohistochemical staining for RPN2 proteins in representative tumors of A549-luc-C8 xenograft lung cancer models. The scale bars indicate 100 μm. (b) Phase-contrast micrographs of A549-luc-C8 cells 96 h after transfection with RPN2 siRNA, RPN2 PnkRNA, RPN2 nkRNA or control siRNA using <t>DharmaFECT</t> 1 reagent. The scale bars indicate 10 μm. (c) Cell proliferation was measured 96 h after transfection with each of the RNAi therapeutic agents. Inhibition of cell growth was observed on A549-luc-C8 cells treated with RPN2 siRNA, PnkRNA, nkRNA, or the control siRNA. Statistical analysis was performed by the Bonferroni multiple-comparison test. The data represent the means ± SD ( n = 3). ***, P
    Dharmafect 1 Reagent, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 304 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Horizon Discovery dharmafect 1 hela
    Suppressive effect of novel RNAi agents for RPN2 in A549-luc-C8 cells. (a) Immunohistochemical staining for RPN2 proteins in representative tumors of A549-luc-C8 xenograft lung cancer models. The scale bars indicate 100 μm. (b) Phase-contrast micrographs of A549-luc-C8 cells 96 h after transfection with RPN2 siRNA, RPN2 PnkRNA, RPN2 nkRNA or control siRNA using <t>DharmaFECT</t> 1 reagent. The scale bars indicate 10 μm. (c) Cell proliferation was measured 96 h after transfection with each of the RNAi therapeutic agents. Inhibition of cell growth was observed on A549-luc-C8 cells treated with RPN2 siRNA, PnkRNA, nkRNA, or the control siRNA. Statistical analysis was performed by the Bonferroni multiple-comparison test. The data represent the means ± SD ( n = 3). ***, P
    Dharmafect 1 Hela, supplied by Horizon Discovery, used in various techniques. Bioz Stars score: 93/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Horizon Discovery dharmafect 1 dharmacon
    Suppressive effect of novel RNAi agents for RPN2 in A549-luc-C8 cells. (a) Immunohistochemical staining for RPN2 proteins in representative tumors of A549-luc-C8 xenograft lung cancer models. The scale bars indicate 100 μm. (b) Phase-contrast micrographs of A549-luc-C8 cells 96 h after transfection with RPN2 siRNA, RPN2 PnkRNA, RPN2 nkRNA or control siRNA using <t>DharmaFECT</t> 1 reagent. The scale bars indicate 10 μm. (c) Cell proliferation was measured 96 h after transfection with each of the RNAi therapeutic agents. Inhibition of cell growth was observed on A549-luc-C8 cells treated with RPN2 siRNA, PnkRNA, nkRNA, or the control siRNA. Statistical analysis was performed by the Bonferroni multiple-comparison test. The data represent the means ± SD ( n = 3). ***, P
    Dharmafect 1 Dharmacon, supplied by Horizon Discovery, used in various techniques. Bioz Stars score: 85/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Horizon Discovery lipid dharmafect 1
    Suppressive effect of novel RNAi agents for RPN2 in A549-luc-C8 cells. (a) Immunohistochemical staining for RPN2 proteins in representative tumors of A549-luc-C8 xenograft lung cancer models. The scale bars indicate 100 μm. (b) Phase-contrast micrographs of A549-luc-C8 cells 96 h after transfection with RPN2 siRNA, RPN2 PnkRNA, RPN2 nkRNA or control siRNA using <t>DharmaFECT</t> 1 reagent. The scale bars indicate 10 μm. (c) Cell proliferation was measured 96 h after transfection with each of the RNAi therapeutic agents. Inhibition of cell growth was observed on A549-luc-C8 cells treated with RPN2 siRNA, PnkRNA, nkRNA, or the control siRNA. Statistical analysis was performed by the Bonferroni multiple-comparison test. The data represent the means ± SD ( n = 3). ***, P
    Lipid Dharmafect 1, supplied by Horizon Discovery, used in various techniques. Bioz Stars score: 90/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher dharmafect 1 kit
    Suppressive effect of novel RNAi agents for RPN2 in A549-luc-C8 cells. (a) Immunohistochemical staining for RPN2 proteins in representative tumors of A549-luc-C8 xenograft lung cancer models. The scale bars indicate 100 μm. (b) Phase-contrast micrographs of A549-luc-C8 cells 96 h after transfection with RPN2 siRNA, RPN2 PnkRNA, RPN2 nkRNA or control siRNA using <t>DharmaFECT</t> 1 reagent. The scale bars indicate 10 μm. (c) Cell proliferation was measured 96 h after transfection with each of the RNAi therapeutic agents. Inhibition of cell growth was observed on A549-luc-C8 cells treated with RPN2 siRNA, PnkRNA, nkRNA, or the control siRNA. Statistical analysis was performed by the Bonferroni multiple-comparison test. The data represent the means ± SD ( n = 3). ***, P
    Dharmafect 1 Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher lipid reagent dharmafect 1
    Suppressive effect of novel RNAi agents for RPN2 in A549-luc-C8 cells. (a) Immunohistochemical staining for RPN2 proteins in representative tumors of A549-luc-C8 xenograft lung cancer models. The scale bars indicate 100 μm. (b) Phase-contrast micrographs of A549-luc-C8 cells 96 h after transfection with RPN2 siRNA, RPN2 PnkRNA, RPN2 nkRNA or control siRNA using <t>DharmaFECT</t> 1 reagent. The scale bars indicate 10 μm. (c) Cell proliferation was measured 96 h after transfection with each of the RNAi therapeutic agents. Inhibition of cell growth was observed on A549-luc-C8 cells treated with RPN2 siRNA, PnkRNA, nkRNA, or the control siRNA. Statistical analysis was performed by the Bonferroni multiple-comparison test. The data represent the means ± SD ( n = 3). ***, P
    Lipid Reagent Dharmafect 1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 89/100, based on 30 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Horizon Discovery transfection dharmafect 1
    Suppressive effect of novel RNAi agents for RPN2 in A549-luc-C8 cells. (a) Immunohistochemical staining for RPN2 proteins in representative tumors of A549-luc-C8 xenograft lung cancer models. The scale bars indicate 100 μm. (b) Phase-contrast micrographs of A549-luc-C8 cells 96 h after transfection with RPN2 siRNA, RPN2 PnkRNA, RPN2 nkRNA or control siRNA using <t>DharmaFECT</t> 1 reagent. The scale bars indicate 10 μm. (c) Cell proliferation was measured 96 h after transfection with each of the RNAi therapeutic agents. Inhibition of cell growth was observed on A549-luc-C8 cells treated with RPN2 siRNA, PnkRNA, nkRNA, or the control siRNA. Statistical analysis was performed by the Bonferroni multiple-comparison test. The data represent the means ± SD ( n = 3). ***, P
    Transfection Dharmafect 1, supplied by Horizon Discovery, used in various techniques. Bioz Stars score: 91/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GE Healthcare dharmafect 1 solution
    Suppressive effect of novel RNAi agents for RPN2 in A549-luc-C8 cells. (a) Immunohistochemical staining for RPN2 proteins in representative tumors of A549-luc-C8 xenograft lung cancer models. The scale bars indicate 100 μm. (b) Phase-contrast micrographs of A549-luc-C8 cells 96 h after transfection with RPN2 siRNA, RPN2 PnkRNA, RPN2 nkRNA or control siRNA using <t>DharmaFECT</t> 1 reagent. The scale bars indicate 10 μm. (c) Cell proliferation was measured 96 h after transfection with each of the RNAi therapeutic agents. Inhibition of cell growth was observed on A549-luc-C8 cells treated with RPN2 siRNA, PnkRNA, nkRNA, or the control siRNA. Statistical analysis was performed by the Bonferroni multiple-comparison test. The data represent the means ± SD ( n = 3). ***, P
    Dharmafect 1 Solution, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 91/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Horizon Discovery bt 474 dharmafect 1
    Suppressive effect of novel RNAi agents for RPN2 in A549-luc-C8 cells. (a) Immunohistochemical staining for RPN2 proteins in representative tumors of A549-luc-C8 xenograft lung cancer models. The scale bars indicate 100 μm. (b) Phase-contrast micrographs of A549-luc-C8 cells 96 h after transfection with RPN2 siRNA, RPN2 PnkRNA, RPN2 nkRNA or control siRNA using <t>DharmaFECT</t> 1 reagent. The scale bars indicate 10 μm. (c) Cell proliferation was measured 96 h after transfection with each of the RNAi therapeutic agents. Inhibition of cell growth was observed on A549-luc-C8 cells treated with RPN2 siRNA, PnkRNA, nkRNA, or the control siRNA. Statistical analysis was performed by the Bonferroni multiple-comparison test. The data represent the means ± SD ( n = 3). ***, P
    Bt 474 Dharmafect 1, supplied by Horizon Discovery, used in various techniques. Bioz Stars score: 93/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Polypus Transfection dharmafect 1
    Suppressive effect of novel RNAi agents for RPN2 in A549-luc-C8 cells. (a) Immunohistochemical staining for RPN2 proteins in representative tumors of A549-luc-C8 xenograft lung cancer models. The scale bars indicate 100 μm. (b) Phase-contrast micrographs of A549-luc-C8 cells 96 h after transfection with RPN2 siRNA, RPN2 PnkRNA, RPN2 nkRNA or control siRNA using <t>DharmaFECT</t> 1 reagent. The scale bars indicate 10 μm. (c) Cell proliferation was measured 96 h after transfection with each of the RNAi therapeutic agents. Inhibition of cell growth was observed on A549-luc-C8 cells treated with RPN2 siRNA, PnkRNA, nkRNA, or the control siRNA. Statistical analysis was performed by the Bonferroni multiple-comparison test. The data represent the means ± SD ( n = 3). ***, P
    Dharmafect 1, supplied by Polypus Transfection, used in various techniques. Bioz Stars score: 93/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher transfection agent dharmafect 1
    Suppressive effect of novel RNAi agents for RPN2 in A549-luc-C8 cells. (a) Immunohistochemical staining for RPN2 proteins in representative tumors of A549-luc-C8 xenograft lung cancer models. The scale bars indicate 100 μm. (b) Phase-contrast micrographs of A549-luc-C8 cells 96 h after transfection with RPN2 siRNA, RPN2 PnkRNA, RPN2 nkRNA or control siRNA using <t>DharmaFECT</t> 1 reagent. The scale bars indicate 10 μm. (c) Cell proliferation was measured 96 h after transfection with each of the RNAi therapeutic agents. Inhibition of cell growth was observed on A549-luc-C8 cells treated with RPN2 siRNA, PnkRNA, nkRNA, or the control siRNA. Statistical analysis was performed by the Bonferroni multiple-comparison test. The data represent the means ± SD ( n = 3). ***, P
    Transfection Agent Dharmafect 1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Horizon Discovery dharmafect 1 transfection reagent
    GLI1 regulates VEGFR2 expression. ( A ) Western blot analysis of protein expression in MDA-MB-468 and HUVEC cells treated with NVP-LDE225 (2.5 μ M ). ( B ) Relative expression of sVEGFR2 mRNA in MDA-MB-468 and HUVEC cells treated with NVP-LDE225 (2.5 μ M ), as performed by real-time RT–PCR (qRT-PCR) analysis. Data were calculated with mean cycle threshold (CT) values, normalised to endogenous control. Data represent the mean (±s.d.) of three independent experiments, each performed in triplicate. ( C ) Western blot analysis of protein expression in MDA-MB-468 and HUVEC cells, 24 and 48 h after <t>transfection</t> with scramble or GLI1 siRNA pool (50 nmol l −1 ) using <t>DharmaFECT</t> 1 Transfection Reagent in DMEM. ( D ) Western blot analysis of protein expression in MDA-MB-231 cells, 24 h after transfection with either pCMV6-GFP empty vector, pCMV6-GFP GLI1 or pCMV6-GFP tGLI1 plasmids using lipofectamine 2000 in DMEM. ( E ) Relative luciferase units in MDA-MB-468 cells transfected with the empty pGL4 plasmid or the pGL4 plasmid containing 500 bp fragment of VEGFR2 promoter, and treated with NVP-LDE225 5 μ M for 24 h after transfection. Luciferase activity was determined 48 h after transfection. Results were the average of three independent experiments. ( F ) Chromatin immunoprecipitation (ChIP) assay in MDA-MB-468 cells by using a GLI1 antibody and primers specific for the VEGFR2 promoter. Results were reported as fold change compared to negative control (no antibody); results were the average of three independent experiments. Bars, s.d. Asterisks indicate statistical significance, as determined by the Student t -test (** P
    Dharmafect 1 Transfection Reagent, supplied by Horizon Discovery, used in various techniques. Bioz Stars score: 92/100, based on 1649 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher dharmafect 1 transfection reagent
    Selection of optimal HER2 siRNA sequences based on the level of HER2 mRNA knockdown, cell viability reduction, number of off-targets and specificity A. The average expression of HER2 mRNA in 3 HER2-positive cell lines (BT474, SKBR3 and HCC1954), measured by QuantiGene assay at 5 days <t>post-transfection</t> (normalized to actin and reported as the percentage of the average of 4 control siRNAs). B. The cell viability reduction induced by the top 10 candidates from (A), measured with CellTiter-Glo assay at 5 days post-transfection. Graph shows the average value from 5 HER2-positive cell lines (BT474, SKBR3, HCC1954, HCC1569, and JIMT1). C. The number of off-targets found through BLAST for the top 5 sequences from (B). D. Cell viability reduction induced by the best 2 sequences from (C) in 18 HER2-positive cell lines (see Supplementary Figure S1 ). E. Cell viability reduction induced by the best 2 sequences in 2 HER2-negative cell lines (T47D and MCF10A). All with 10 nM of siRNA delivered with <t>DharmaFECT-1.</t>
    Dharmafect 1 Transfection Reagent, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 1396 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    ( a ) A stably transfected vector in HeLa cells expresses two luciferase genes. Firefly luciferase is expressed constitutively, whereas expression of the renilla luciferase gene is under the control of miR-15b as part of the RNA-induced silencing complex (RISC). In the presence of an anti-miR-15b oligonucleotide (TP ODN in this figure), the RISC dissociates from the renilla gene. This leads to an increase in renilla luciferase activity. Comparison of the activity of renilla luciferase to the unregulated firefly luciferase yields an indication of the activity of an oligonucleotide as an anti-miR-15b oligonucleotide. ( a ) Adapted from the Integrated DNA Technologies Inc. (Coralville, IA, USA) website. The stably transfected HeLa cell line and assay protocols were developed by miRagen Therapeutics, Inc. ( b ) Lipid transfection using DharmaFECT 1 (Dharmacon). ( c ) Passive transfection using TP ODNs. Positive and negative controls were purchased from Dharmacon: miRIDIAN Hairpin Inhibitor (IH-300587-07) and miRIDIAN Hairpin Inhibitor Negative Control #1 (IN-001005-01).

    Journal: Signal Transduction and Targeted Therapy

    Article Title: Peptide-substituted oligonucleotide synthesis and non-toxic, passive cell delivery

    doi: 10.1038/sigtrans.2016.19

    Figure Lengend Snippet: ( a ) A stably transfected vector in HeLa cells expresses two luciferase genes. Firefly luciferase is expressed constitutively, whereas expression of the renilla luciferase gene is under the control of miR-15b as part of the RNA-induced silencing complex (RISC). In the presence of an anti-miR-15b oligonucleotide (TP ODN in this figure), the RISC dissociates from the renilla gene. This leads to an increase in renilla luciferase activity. Comparison of the activity of renilla luciferase to the unregulated firefly luciferase yields an indication of the activity of an oligonucleotide as an anti-miR-15b oligonucleotide. ( a ) Adapted from the Integrated DNA Technologies Inc. (Coralville, IA, USA) website. The stably transfected HeLa cell line and assay protocols were developed by miRagen Therapeutics, Inc. ( b ) Lipid transfection using DharmaFECT 1 (Dharmacon). ( c ) Passive transfection using TP ODNs. Positive and negative controls were purchased from Dharmacon: miRIDIAN Hairpin Inhibitor (IH-300587-07) and miRIDIAN Hairpin Inhibitor Negative Control #1 (IN-001005-01).

    Article Snippet: TP ODN and DharmaFECT 1 (Dharmacon) were mixed and allowed to form a complex at room temperature (20 min).

    Techniques: Stable Transfection, Transfection, Plasmid Preparation, Luciferase, Expressing, Activity Assay, IA, Negative Control

    Localization of fluorescein-labeled ODNs in HeLa cells. ( a ) Lipid transfection: 100 nM fluorescein-labeled 4K (left) and 6K (right) were transfected into HeLa cells using DharmaFECT 1 (Dharmacon) reagent for 18 h followed by washing. ( b ) Passive transfection: HeLa cells were incubated with 1 μM fluorescein-labeled 4K (left) and 6K (right) for 90 h followed by washing. Cells were stained with Hoechst 33258 to visualize the nuclei.

    Journal: Signal Transduction and Targeted Therapy

    Article Title: Peptide-substituted oligonucleotide synthesis and non-toxic, passive cell delivery

    doi: 10.1038/sigtrans.2016.19

    Figure Lengend Snippet: Localization of fluorescein-labeled ODNs in HeLa cells. ( a ) Lipid transfection: 100 nM fluorescein-labeled 4K (left) and 6K (right) were transfected into HeLa cells using DharmaFECT 1 (Dharmacon) reagent for 18 h followed by washing. ( b ) Passive transfection: HeLa cells were incubated with 1 μM fluorescein-labeled 4K (left) and 6K (right) for 90 h followed by washing. Cells were stained with Hoechst 33258 to visualize the nuclei.

    Article Snippet: TP ODN and DharmaFECT 1 (Dharmacon) were mixed and allowed to form a complex at room temperature (20 min).

    Techniques: Labeling, Transfection, Incubation, Staining

    Effect of VEGF and VEGF Receptor siRNA Oligonucleotides on the Survival of MCF-7 Cells (A) MCF-7 cells were transiently transfected with various VEGF or VEGF receptor siRNA oligonucleotides, as indicated. Transfection procedures were performed with DharmaFECT-1 reagent according to the manufacturer's protocol. After 5 d of incubation, the cells were lysed and subjected to Western blotting (WB) using anti-VEGFR1 or anti-NRP1 antibodies. Total protein extracts were analyzed by Western blotting using anti-Csk antibody as an internal control. Alternatively, after 5 d of transfection, total RNA (5 μg) was isolated and subjected to RT-PCR analysis for VEGF and VEGFR2 mRNA expression. GAPDH mRNA is shown as an internal control. (B) MCF-7 cells were transiently transfected with siLuc, siNRP1, siVEGF, or various VEGF receptor siRNA oligonucleotides in the absence or presence of PGF (20 ng/ml) or VEGF (20 ng/ml), as indicated. After 5 d of incubation, the cells were harvested and subjected to cell cycle analysis. The data are representative of three individual studies. m, mutant; w, wild type. (C) MCF-7 cells were transiently transfected with siLuc, siNRP1, siVEGF, or various VEGF receptor siRNA oligonucleotides as shown. After 5 d of incubation, the cells were observed under a light microscope. (D) Effect of VEGF on the cell cycle in MDA-MB-231 cells. MDA-MB-231 cells were transiently transfected with siLuc or with siVEGF oligonucleotides in the presence or absence of VEGF (20 ng/ml). After 5 d of incubation, cells were harvested and subjected to cell cycle analysis. The data are representative of three individual studies.

    Journal: PLoS Medicine

    Article Title: Vascular Endothelial Growth Factor Mediates Intracrine Survival in Human Breast Carcinoma Cells through Internally Expressed VEGFR1/FLT1

    doi: 10.1371/journal.pmed.0040186

    Figure Lengend Snippet: Effect of VEGF and VEGF Receptor siRNA Oligonucleotides on the Survival of MCF-7 Cells (A) MCF-7 cells were transiently transfected with various VEGF or VEGF receptor siRNA oligonucleotides, as indicated. Transfection procedures were performed with DharmaFECT-1 reagent according to the manufacturer's protocol. After 5 d of incubation, the cells were lysed and subjected to Western blotting (WB) using anti-VEGFR1 or anti-NRP1 antibodies. Total protein extracts were analyzed by Western blotting using anti-Csk antibody as an internal control. Alternatively, after 5 d of transfection, total RNA (5 μg) was isolated and subjected to RT-PCR analysis for VEGF and VEGFR2 mRNA expression. GAPDH mRNA is shown as an internal control. (B) MCF-7 cells were transiently transfected with siLuc, siNRP1, siVEGF, or various VEGF receptor siRNA oligonucleotides in the absence or presence of PGF (20 ng/ml) or VEGF (20 ng/ml), as indicated. After 5 d of incubation, the cells were harvested and subjected to cell cycle analysis. The data are representative of three individual studies. m, mutant; w, wild type. (C) MCF-7 cells were transiently transfected with siLuc, siNRP1, siVEGF, or various VEGF receptor siRNA oligonucleotides as shown. After 5 d of incubation, the cells were observed under a light microscope. (D) Effect of VEGF on the cell cycle in MDA-MB-231 cells. MDA-MB-231 cells were transiently transfected with siLuc or with siVEGF oligonucleotides in the presence or absence of VEGF (20 ng/ml). After 5 d of incubation, cells were harvested and subjected to cell cycle analysis. The data are representative of three individual studies.

    Article Snippet: Transfection procedures were performed with Oligofectamine Reagent (Invitrogen) in MDA-MB-231 cells or with DharmaFECT-1 reagent (Dharmacon) in MCF-7 cells according to the manufacturers' protocols.

    Techniques: Transfection, Incubation, Western Blot, Isolation, Reverse Transcription Polymerase Chain Reaction, Expressing, Cell Cycle Assay, Mutagenesis, Light Microscopy, Multiple Displacement Amplification

    ( a ) A stably transfected vector in HeLa cells expresses two luciferase genes. Firefly luciferase is expressed constitutively, whereas expression of the renilla luciferase gene is under the control of miR-15b as part of the RNA-induced silencing complex (RISC). In the presence of an anti-miR-15b oligonucleotide (TP ODN in this figure), the RISC dissociates from the renilla gene. This leads to an increase in renilla luciferase activity. Comparison of the activity of renilla luciferase to the unregulated firefly luciferase yields an indication of the activity of an oligonucleotide as an anti-miR-15b oligonucleotide. ( a ) Adapted from the Integrated DNA Technologies Inc. (Coralville, IA, USA) website. The stably transfected HeLa cell line and assay protocols were developed by miRagen Therapeutics, Inc. ( b ) Lipid transfection using DharmaFECT 1 (Dharmacon). ( c ) Passive transfection using TP ODNs. Positive and negative controls were purchased from Dharmacon: miRIDIAN Hairpin Inhibitor (IH-300587-07) and miRIDIAN Hairpin Inhibitor Negative Control #1 (IN-001005-01).

    Journal: Signal Transduction and Targeted Therapy

    Article Title: Peptide-substituted oligonucleotide synthesis and non-toxic, passive cell delivery

    doi: 10.1038/sigtrans.2016.19

    Figure Lengend Snippet: ( a ) A stably transfected vector in HeLa cells expresses two luciferase genes. Firefly luciferase is expressed constitutively, whereas expression of the renilla luciferase gene is under the control of miR-15b as part of the RNA-induced silencing complex (RISC). In the presence of an anti-miR-15b oligonucleotide (TP ODN in this figure), the RISC dissociates from the renilla gene. This leads to an increase in renilla luciferase activity. Comparison of the activity of renilla luciferase to the unregulated firefly luciferase yields an indication of the activity of an oligonucleotide as an anti-miR-15b oligonucleotide. ( a ) Adapted from the Integrated DNA Technologies Inc. (Coralville, IA, USA) website. The stably transfected HeLa cell line and assay protocols were developed by miRagen Therapeutics, Inc. ( b ) Lipid transfection using DharmaFECT 1 (Dharmacon). ( c ) Passive transfection using TP ODNs. Positive and negative controls were purchased from Dharmacon: miRIDIAN Hairpin Inhibitor (IH-300587-07) and miRIDIAN Hairpin Inhibitor Negative Control #1 (IN-001005-01).

    Article Snippet: In an eppendorf tube, 0.25 μl DharmaFECT 1 (purchased from Dharmacon/ThermoFisher, now GE Dharmacon, Lafayette, CO, USA) was added to 50 μl of the reduced serum medium and this solution was added to the TP ODN in the same medium.

    Techniques: Stable Transfection, Transfection, Plasmid Preparation, Luciferase, Expressing, Activity Assay, IA, Negative Control

    Localization of fluorescein-labeled ODNs in HeLa cells. ( a ) Lipid transfection: 100 nM fluorescein-labeled 4K (left) and 6K (right) were transfected into HeLa cells using DharmaFECT 1 (Dharmacon) reagent for 18 h followed by washing. ( b ) Passive transfection: HeLa cells were incubated with 1 μM fluorescein-labeled 4K (left) and 6K (right) for 90 h followed by washing. Cells were stained with Hoechst 33258 to visualize the nuclei.

    Journal: Signal Transduction and Targeted Therapy

    Article Title: Peptide-substituted oligonucleotide synthesis and non-toxic, passive cell delivery

    doi: 10.1038/sigtrans.2016.19

    Figure Lengend Snippet: Localization of fluorescein-labeled ODNs in HeLa cells. ( a ) Lipid transfection: 100 nM fluorescein-labeled 4K (left) and 6K (right) were transfected into HeLa cells using DharmaFECT 1 (Dharmacon) reagent for 18 h followed by washing. ( b ) Passive transfection: HeLa cells were incubated with 1 μM fluorescein-labeled 4K (left) and 6K (right) for 90 h followed by washing. Cells were stained with Hoechst 33258 to visualize the nuclei.

    Article Snippet: In an eppendorf tube, 0.25 μl DharmaFECT 1 (purchased from Dharmacon/ThermoFisher, now GE Dharmacon, Lafayette, CO, USA) was added to 50 μl of the reduced serum medium and this solution was added to the TP ODN in the same medium.

    Techniques: Labeling, Transfection, Incubation, Staining

    Suppressive effect of novel RNAi agents for RPN2 in A549-luc-C8 cells. (a) Immunohistochemical staining for RPN2 proteins in representative tumors of A549-luc-C8 xenograft lung cancer models. The scale bars indicate 100 μm. (b) Phase-contrast micrographs of A549-luc-C8 cells 96 h after transfection with RPN2 siRNA, RPN2 PnkRNA, RPN2 nkRNA or control siRNA using DharmaFECT 1 reagent. The scale bars indicate 10 μm. (c) Cell proliferation was measured 96 h after transfection with each of the RNAi therapeutic agents. Inhibition of cell growth was observed on A549-luc-C8 cells treated with RPN2 siRNA, PnkRNA, nkRNA, or the control siRNA. Statistical analysis was performed by the Bonferroni multiple-comparison test. The data represent the means ± SD ( n = 3). ***, P

    Journal: Scientific Reports

    Article Title: A novel platform to enable inhaled naked RNAi medicine for lung cancer

    doi: 10.1038/srep03325

    Figure Lengend Snippet: Suppressive effect of novel RNAi agents for RPN2 in A549-luc-C8 cells. (a) Immunohistochemical staining for RPN2 proteins in representative tumors of A549-luc-C8 xenograft lung cancer models. The scale bars indicate 100 μm. (b) Phase-contrast micrographs of A549-luc-C8 cells 96 h after transfection with RPN2 siRNA, RPN2 PnkRNA, RPN2 nkRNA or control siRNA using DharmaFECT 1 reagent. The scale bars indicate 10 μm. (c) Cell proliferation was measured 96 h after transfection with each of the RNAi therapeutic agents. Inhibition of cell growth was observed on A549-luc-C8 cells treated with RPN2 siRNA, PnkRNA, nkRNA, or the control siRNA. Statistical analysis was performed by the Bonferroni multiple-comparison test. The data represent the means ± SD ( n = 3). ***, P

    Article Snippet: Each labeled oligonucleotide was transfected with or without DharmaFECT 1 reagent.

    Techniques: Immunohistochemistry, Staining, Transfection, Inhibition

    GLI1 regulates VEGFR2 expression. ( A ) Western blot analysis of protein expression in MDA-MB-468 and HUVEC cells treated with NVP-LDE225 (2.5 μ M ). ( B ) Relative expression of sVEGFR2 mRNA in MDA-MB-468 and HUVEC cells treated with NVP-LDE225 (2.5 μ M ), as performed by real-time RT–PCR (qRT-PCR) analysis. Data were calculated with mean cycle threshold (CT) values, normalised to endogenous control. Data represent the mean (±s.d.) of three independent experiments, each performed in triplicate. ( C ) Western blot analysis of protein expression in MDA-MB-468 and HUVEC cells, 24 and 48 h after transfection with scramble or GLI1 siRNA pool (50 nmol l −1 ) using DharmaFECT 1 Transfection Reagent in DMEM. ( D ) Western blot analysis of protein expression in MDA-MB-231 cells, 24 h after transfection with either pCMV6-GFP empty vector, pCMV6-GFP GLI1 or pCMV6-GFP tGLI1 plasmids using lipofectamine 2000 in DMEM. ( E ) Relative luciferase units in MDA-MB-468 cells transfected with the empty pGL4 plasmid or the pGL4 plasmid containing 500 bp fragment of VEGFR2 promoter, and treated with NVP-LDE225 5 μ M for 24 h after transfection. Luciferase activity was determined 48 h after transfection. Results were the average of three independent experiments. ( F ) Chromatin immunoprecipitation (ChIP) assay in MDA-MB-468 cells by using a GLI1 antibody and primers specific for the VEGFR2 promoter. Results were reported as fold change compared to negative control (no antibody); results were the average of three independent experiments. Bars, s.d. Asterisks indicate statistical significance, as determined by the Student t -test (** P

    Journal: British Journal of Cancer

    Article Title: Hedgehog signalling pathway orchestrates angiogenesis in triple-negative breast cancers

    doi: 10.1038/bjc.2017.116

    Figure Lengend Snippet: GLI1 regulates VEGFR2 expression. ( A ) Western blot analysis of protein expression in MDA-MB-468 and HUVEC cells treated with NVP-LDE225 (2.5 μ M ). ( B ) Relative expression of sVEGFR2 mRNA in MDA-MB-468 and HUVEC cells treated with NVP-LDE225 (2.5 μ M ), as performed by real-time RT–PCR (qRT-PCR) analysis. Data were calculated with mean cycle threshold (CT) values, normalised to endogenous control. Data represent the mean (±s.d.) of three independent experiments, each performed in triplicate. ( C ) Western blot analysis of protein expression in MDA-MB-468 and HUVEC cells, 24 and 48 h after transfection with scramble or GLI1 siRNA pool (50 nmol l −1 ) using DharmaFECT 1 Transfection Reagent in DMEM. ( D ) Western blot analysis of protein expression in MDA-MB-231 cells, 24 h after transfection with either pCMV6-GFP empty vector, pCMV6-GFP GLI1 or pCMV6-GFP tGLI1 plasmids using lipofectamine 2000 in DMEM. ( E ) Relative luciferase units in MDA-MB-468 cells transfected with the empty pGL4 plasmid or the pGL4 plasmid containing 500 bp fragment of VEGFR2 promoter, and treated with NVP-LDE225 5 μ M for 24 h after transfection. Luciferase activity was determined 48 h after transfection. Results were the average of three independent experiments. ( F ) Chromatin immunoprecipitation (ChIP) assay in MDA-MB-468 cells by using a GLI1 antibody and primers specific for the VEGFR2 promoter. Results were reported as fold change compared to negative control (no antibody); results were the average of three independent experiments. Bars, s.d. Asterisks indicate statistical significance, as determined by the Student t -test (** P

    Article Snippet: For siRNA validation, cells were transfected with GLI1 siRNAs (50 nmol l−1 ) using DharmaFECT 1 Transfection Reagent in DMEM (GE Dharmacon); 24 and 48 h after transfection, western blot analysis for GLI1 and VEGFR2 expression was performed.

    Techniques: Expressing, Western Blot, Multiple Displacement Amplification, Quantitative RT-PCR, Transfection, Plasmid Preparation, Luciferase, Activity Assay, Chromatin Immunoprecipitation, Negative Control

    Selection of optimal HER2 siRNA sequences based on the level of HER2 mRNA knockdown, cell viability reduction, number of off-targets and specificity A. The average expression of HER2 mRNA in 3 HER2-positive cell lines (BT474, SKBR3 and HCC1954), measured by QuantiGene assay at 5 days post-transfection (normalized to actin and reported as the percentage of the average of 4 control siRNAs). B. The cell viability reduction induced by the top 10 candidates from (A), measured with CellTiter-Glo assay at 5 days post-transfection. Graph shows the average value from 5 HER2-positive cell lines (BT474, SKBR3, HCC1954, HCC1569, and JIMT1). C. The number of off-targets found through BLAST for the top 5 sequences from (B). D. Cell viability reduction induced by the best 2 sequences from (C) in 18 HER2-positive cell lines (see Supplementary Figure S1 ). E. Cell viability reduction induced by the best 2 sequences in 2 HER2-negative cell lines (T47D and MCF10A). All with 10 nM of siRNA delivered with DharmaFECT-1.

    Journal: Oncotarget

    Article Title: Therapeutic siRNA for drug-resistant HER2-positive breast cancer

    doi: 10.18632/oncotarget.7409

    Figure Lengend Snippet: Selection of optimal HER2 siRNA sequences based on the level of HER2 mRNA knockdown, cell viability reduction, number of off-targets and specificity A. The average expression of HER2 mRNA in 3 HER2-positive cell lines (BT474, SKBR3 and HCC1954), measured by QuantiGene assay at 5 days post-transfection (normalized to actin and reported as the percentage of the average of 4 control siRNAs). B. The cell viability reduction induced by the top 10 candidates from (A), measured with CellTiter-Glo assay at 5 days post-transfection. Graph shows the average value from 5 HER2-positive cell lines (BT474, SKBR3, HCC1954, HCC1569, and JIMT1). C. The number of off-targets found through BLAST for the top 5 sequences from (B). D. Cell viability reduction induced by the best 2 sequences from (C) in 18 HER2-positive cell lines (see Supplementary Figure S1 ). E. Cell viability reduction induced by the best 2 sequences in 2 HER2-negative cell lines (T47D and MCF10A). All with 10 nM of siRNA delivered with DharmaFECT-1.

    Article Snippet: Transfection was carried out using DharmaFECT-1 transfection reagent (Thermo Scientific Dharmacon) diluted in OptiMEM medium (Life Technologies).

    Techniques: Selection, Expressing, Transfection, Glo Assay